One form of tissue analysis comprises staining tissue with a dye and placing this stained tissue on a microscope slide. The tissue may be stained with more than one dye for a number of different reasons. For example, two dyes may be used to identify the presence of two different proteins. In another application, one dye may be specific to bind with a particular cell component such as cytoplasm, while the other dye may stain nuclei. The microscope slide is then illuminated and an electronic image is produced. A digital video image of stained tissue may be produced. Pixels in the image have color components. In order to derive separate signals, each indicative of level of a different color in the image, the video signal must be “unmixed.” Various statistical methods may be used to process the color components to obtain indications of color in the image. These indications must then be further processed to provide information of interest. Manual processing of the information has been provided by having a pathologist review and evaluate the slide. Automated analysis has also been provided for certain forms of slide evaluation but not others.
U.S. Pat. No. 6,631,203 shows a charge coupled device (CCD) camera providing an image of a stained slide to an image processor.
U.S. Pat. No. 5,995,645 discloses a method for cancer cell detection including the steps of staining an analyzed sample with at least first and second dyes, the dyes being selected such that the first dye better adheres to normal cells whereas the second dye better adheres to cancer cells; spectrally imaging the sample through an optical device being optically connected to an imaging spectrometer, thereby obtaining a spectrum of each pixel of the sample; based on the spectra, evaluating concentrations of the first and second dyes for each of the pixels in an image; and based on the concentrations detecting the presence of cancer cells in the sample.
U.S. Pat. No. 6,665,438 discloses unmixing hyperspectral imagery data using a genetic algorithm and robust Kalman filtering techniques, in a pixel by pixel fashion, until a hyperspectral image cube is completed.
U.S. Pat. No. 6,728,396 relates generally to statistical signal processing and its application to imaging, and more particularly to a method and/or associated apparatus for independent component imaging from mixed observations. The statistical and/or neural computation technique of Independent Component Analysis (ICA) is used for separating hidden sources from mixed image signals.
U.S. Pat. No. 6,697,509 discloses an imager used to generate an image of a sample, e.g., a tissue sample including diaminobenzidine tetrahydrochloride (DAB) stained cells. A user selects a region to be scored in the image. A mean intensity value of a selected color is determined from the pixels in the selected region. The selected color may be the complement of a color to be scored, which, in the case of a DAB test, is blue. A score is generated in response to the mean intensity value using a stored calibration curve.
U.S. Pat. No. 6,675,106 discloses a method of determining the properties of a sample from measured spectral data collected from the sample by performing a multivariate spectral analysis. The method can include generating a two-dimensional matrix A containing measured spectral data; providing a weighted spectral data matrix D by performing a weighting operation on matrix A; factoring D into the product of two matrices C and ST, by performing a constrained alternating least-squares analysis of D=CST, where C is a concentration intensity matrix and S is a spectral shapes matrix; unweighting C and S by applying the inverse of the weighting used previously; and determining the properties of the sample by inspecting C and S.
U.S. Pat. No. 6,427,126 discusses an advanced imaging spectrograph system and method for very high throughput identification, sequencing and/or genotyping of DNA and other molecules. The system is based on the integration of improved electrophoresis structures with an imaging spectrophotometer that records the entire emission spectra along an imaging line across a sequencing gel (or capillary array). The system includes spectral shape matching to improve dye identification allowing the u se of dyes having nearly any emission spectra and allowing greater than four-dye multiplexing.
U.S. Pat. No. 6,151,414 discloses a code utilizing an n×p signal matrix which is factored into matrices whose entries are non-negative.
There is nowhere disclosed in the known prior art an arrangement directed to automatic color unmixing in an image of a slide having tissue stained by a first, non-specific dye and by a second, immunohistochemical dye having a color differing from that of the first dye. The immunohistochemical dye could, for example, be specific to cancer cells. The prior art further does not disclose analyzing the image in the spectrum of the immunohistochemical dye for areas of interest, e.g. organs as opposed to stroma (for example fatty or other intermediate tissue). Further, the prior art does not disclose normalizing the color concentration of the second dye with respect to an area of interest to generate a color index having diagnostic significance.